CN103468577B - Nannochloropsis sp mutant strain and application thereof - Google Patents
Nannochloropsis sp mutant strain and application thereof Download PDFInfo
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- CN103468577B CN103468577B CN201310433259.0A CN201310433259A CN103468577B CN 103468577 B CN103468577 B CN 103468577B CN 201310433259 A CN201310433259 A CN 201310433259A CN 103468577 B CN103468577 B CN 103468577B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The invention discloses Nannochloropsis sp and an application thereof. The Nannochloropsis sp disclosed by the invention has the advantages of strong environmental adaptability, strong high temperature tolerance, high content of grease, especially eicosapentaenoic acid, strong CO2 treatment capacity, and better suitability for industrial application and the like, and can be used for production of biodiesel, production of foods, health care products, feeds and baits for aquatic products and production of medicaments, as well as high-temperature farming.
Description
Technical field
The present invention relates to microorganism field, particularly relate to a kind of plan Nannochloropsis oceanica mutant strain ENN11-3 (Nannochloropsis sp) and application thereof.
Background technology
Along with the progressively exhaustion of world's fossil fuel capacity, the continuous increase of CO2 emissions and global climate day by day become warm, the survival and development situation of the mankind has faced unprecedented challenge.Therefore; therefore, countries in the world all actively find can the renewable energy source of petroleum replacing product, wherein; biofuel is exactly a kind of important bioenergy; but such as the cultivation period such as corn and soybean is longer for common oil crops, takies farmland more; " striving grain with people; strive ground with grain " problem can be produced, thus cause the awkward result of " solve energy dilemma, but occur crisis in food ".
Micro-algae be a class on land, ocean is widely distributed, nutritious, growth cycle is short, photosynthetic availability high with the energy-producing lower plant of photoautotrophy, be nature origin the earliest, the widest, the kind of distribution and the maximum biomass resource of quantity.Many micro-algaes can accumulate in a large number shelf stability triacylglycerol (micro-algae oil) (being similar to vegetables oil) in cell under certain condition, and its most high-content can reach more than 50% of dry cell weight.Compared with high Lu Sheng oil crops, the unit surface oil yield of micro-algae is the decades of times of oilseed plant oil yield, can the multiple biofuel such as production biofuel through biological smelting, be considered to the most potential most possible biomass resource becoming oil substitutes.
Except oil yield is very high, micro-algae is rich in proteins, fat, carbohydrate, each seed amino acid, VITAMIN, microbiotic, high unsaturated fatty acid and other various bioactivators also, therefore, in fields such as protective foods, medicine and feeds, there is good DEVELOPMENT PROSPECT too, such as, for foodstuff and pharmaceutical sector (nutritional supplement of humans and animals: VITAMIN, protein, lipid acid, polysaccharide etc.); Extract chemical industry as makeup, fine chemical product etc.; Biogas, fuel is produced as the energy; For bait and feed industry (bait (CN1742570), poultry feed etc. of the aquatic animal such as fishes and shrimps, crustacean); Agriculturally as soil redeposition, fertilizer etc.
Intending Nannochloropsis oceanica (Nannochloropsis sp.) is a kind of important micro-algae of marine products economy, its fast growth, strong stress resistance, and lipid acid composition is simple, and being rich in EPA, is the very potential algae kind of one.The optimum growth temperature intending Nannochloropsis oceanica is 25 DEG C ~ 30 DEG C, and when temperature is more than 35 DEG C, its growth is completely suppressed, and frustule can be dead gradually.China's most area scorching temperature in summer, intends Nannochloropsis oceanica and is difficult to carry out large-scale farming in summer.If by the algae strain of mutagenic obtained withstand high temperatures, will the range of application of this algae be expanded, Nannochloropsis oceanica cultivation is intended to China significant.
Research and the screening operation of current plan Nannochloropsis oceanica launch, Ratnesh Chaturvedi and Yuji Fujita(A Isolation of enhanced eicosapentaenoic acid producing mutants of Nannochloropsis oculata ST-6using ethyl methane sulfonate induced mutagenesis techniques and their characterization at mRNA transcript level, Phycological Research54:208 – 219, 2006.) screen for the fatty acid content intending the strain of Nannochloropsis oceanica algae, but the high temperature tolerance ability of the algae strain screened does not significantly improve, and this algae strain whether to extend to outside scenery also still to be tested.(Temperature stress process improves the fatty acid content change intending intending the strain of Nannochloropsis oceanica algae under microballoon algae (Nannochloropsis oculata) have studied Temperature stress treatment condition to the people such as Yang Guanpin, does not obtain the algae strain of high temperature tolerance capability improving.
Therefore, the plan Nannochloropsis oceanica algae strain with high temperature tolerance ability is needed in the art.
Summary of the invention
The object of the present invention is to provide a kind of strong to adaptive capacity to environment, high temperature tolerance ability is strong, and fat content is high, to CO
2the strong plan Nannochloropsis oceanica being more suitable for industrial application of processing power.
For reaching this object, the present invention by the following technical solutions:
In first aspect, the invention provides a kind of plan Nannochloropsis oceanica ENN11-3 (Nannochloropsis sp.), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation date is on September 5th, 2012, and deposit number is CGMCC No.6446.
The growth tolerance top temperature of plan Nannochloropsis oceanica ENN11-3 provided by the invention is 42 DEG C.。
Plan Nannochloropsis oceanica ENN11-3 provided by the invention can grow at 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C, 30 DEG C, 31 DEG C, 32 DEG C, 33 DEG C, 34 DEG C, 35 DEG C, 36 DEG C, 37 DEG C, 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C.
In second aspect, the invention provides the cultural method of plan Nannochloropsis oceanica ENN11-3 as described in relation to the first aspect, comprise and plan Nannochloropsis oceanica algae strain ENN11-3 green cell is seeded in sea water medium, wherein intensity of illumination 50-500 μm of ol/m
2.s, preferred 100-450 μm ol/m
2.s, more preferably 150-400 μm of ol/m
2.s, also more preferably 200-350 μm of ol/m
2.s, most preferably 250-300 μm of ol/m
2.s; PH value 7-9, preferred 7.5-8.5, more preferably 8; Temperature 25-42 DEG C, preferred 30-38 DEG C; And in incubation period, by passing into the air that carbonic acid gas volume fraction is 1.5-2% in nutrient solution.
In the cultural method of plan Nannochloropsis oceanica ENN11-3 of the present invention, intensity of illumination can be 50 μm of ol/m
2.s, 80 μm of ol/m
2.s, 100 μm of ol/m
2.s, 120 μm of ol/m
2.s, 150 μm of ol/m
2.s, 180 μm of ol/m
2.s, 200 μm of ol/m
2.s, 220 μm of ol/m
2.s, 250 μm of ol/m
2.s, 280 μm of ol/m
2.s, 300 μm of ol/m
2.s, 320 μm of ol/m
2.s, 350 μm of ol/m
2.s, 380 μm of ol/m
2.s, 400 μm of ol/m
2.s, 420 μm of ol/m
2.s, 450 μm of ol/m
2.s, 480 μm of ol/m
2.s or 500 μm of ol/m
2.s; PH value can be 7,7.2,7.5,7.8,8,8.2,8.5,8.8 or 9; Temperature can be 25 DEG C, 26 DEG C, 27 DEG C, 28 DEG C, 29 DEG C, 30 DEG C, 31 DEG C, 32 DEG C, 33 DEG C, 34 DEG C, 35 DEG C, 36 DEG C, 37 DEG C, 38 DEG C, 39 DEG C, 40 DEG C, 41 DEG C or 42 DEG C.
In the third aspect, the invention provides the application of plan Nannochloropsis oceanica ENN11-3 in lipid acid is produced as described in relation to the first aspect, preferably, described lipid acid is capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid, is preferably timnodonic acid.
In fourth aspect, the invention provides the application of plan Nannochloropsis oceanica ENN11-3 in biofuel is produced as described in relation to the first aspect, preferably, described biofuel is biofuel or bio-ethanol.
In the 5th, the invention provides the application of plan Nannochloropsis oceanica ENN11-3 in environment protection as described in relation to the first aspect, preferably described environment protection is CO
2reduce discharging.
In the 6th, the plan Nannochloropsis oceanica ENN11-3 that the invention provides as described in relation to the first aspect is intending the application in the cultivation of Nannochloropsis oceanica high temperature, and wherein said high temperature is 35-42 DEG C.
In the 7th, the invention provides the purposes of plan Nannochloropsis oceanica ENN11-3 in the preparation of the medicine of capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid as described in relation to the first aspect, preferably prepare the application comprised in the medicine of timnodonic acid.
In eighth aspect, the invention provides plan Nannochloropsis oceanica ENN11-3 as described in relation to the first aspect comprise the food of capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid, healthcare products, feed and/or fishery products bait produce in purposes.
In the present invention, sea water medium is algae media the most frequently used in this area, and its concrete component sees below.
Advantageous Effects of the present invention is:
1, by force, high temperature tolerance ability is strong for this algae kind stable performance, environmental compatibility, is applicable to Industry Promotion
2, this algae kind oil and fat accumulation is fast, and oleaginousness is high, is applicable to for production biofuel
3, this algae kind Content of Eicosapentaenoic Acid is higher, is applicable to for producing related drugs, food, healthcare products, feed, fishery products bait etc.
Accompanying drawing explanation
Fig. 1 is the thetagram cultivating plan Nannochloropsis oceanica ENN11-3 of the present invention in indoor column reactor.
Fig. 2 is the dry weight change curve cultivating plan Nannochloropsis oceanica ENN11-3 of the present invention in indoor column reactor.
Fig. 3 is fatty acid content (the accounting for algae powder) variation diagram cultivating plan Nannochloropsis oceanica ENN11-3 of the present invention in indoor column reactor.
Fig. 4 is the temperature variation curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out the cultivation of outdoor tubular type high temperature.
Fig. 5 is the dry weight change curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out the cultivation of outdoor tubular type high temperature.
Fig. 6 is the temperature variation curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out indoor board-like high temperature cultivation.
Fig. 7 is the dry weight change curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out indoor board-like high temperature cultivation.
Fig. 8 is fatty acid content (the accounting for algae powder) figure that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out outdoor board-like high temperature cultivation.
Fig. 9 is the temperature variation curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out outdoor board-like waste gas cultivation.
Figure 10 is the OD750 change curve that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out outdoor board-like waste gas cultivation.
Figure 11 is fatty acid content (the accounting for algae powder) figure that plan Nannochloropsis oceanica ENN11-3 of the present invention carries out outdoor board-like waste gas cultivation.
Detailed Description Of The Invention
Described algae strain ENN11-3 is preserved in (No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms's common micro-organisms center on September 5th, 2012 by contriver, be called for short CGMCC), its preserving number is CGMCC No.6446.
Technical scheme of the present invention is further illustrated by embodiment below in conjunction with accompanying drawing.
The mutagenesis screening of embodiment 1ENN11-3 algae strain, indoor culture and oil and fat accumulation
The fresh sterile algae liquid of plan Nannochloropsis oceanica of phase of taking the logarithm carries out single function alkylating agent N-methyl-N'-nitro-N-nitroso-guani dine (MNNG) mutagenic treatment and outdoor high temperature pressure treatment, lay dull and stereotyped, totally 30 strains that fall of the individual larger single algae of picking enter high temperature primary dcreening operation and high temperature sieves again, obtain the strain of ENN11-3 algae.
Be seeded in the sea water medium (formula is in table 1) prepared by green for the ENN11-3 being in logarithmic phase swarm cell, making cell density reach OD750 is between 0.8-1.2, with wild-type ENN11 for contrast.Culturing process intensity of illumination controls at 50-500umol/m
2.s, in incubation period, by passing into the carbonic acid gas of 1.5-2% and the mixed gas of air in nutrient solution, the pH value of substratum is regulated between 7-9.The column reactor that the reactor that cultivation uses is 40mm internal diameter, length 600mm.Arrangement for heating or cooling water in receptacle for live fish temperature control, room temperature 25 DEG C was cultivated after 4 days, and the 5th day temperature rises to 8 hours daytimes 38 DEG C, night 30 DEG C; From the 6th day, temperature rose to 6 hours daytimes 42 DEG C, all the other times 30 DEG C (temperature variation as shown in Figure 1).In culturing process, timing sampling measures dry weight, the results are shown in Figure 2.Cultivation proceeds to the 15th day, is collected by algae liquid, obtains algae mud, algae mud is carried out vacuum lyophilization again by method that is centrifugal or natural subsidence.
Table 1
Fe-EDTA solution: by 2.76g FeCl
3.6H
2o is dissolved in 1000mL0.05M Na2EDTA solution trace element solution
After drying completes, measure its oil component content as following table 2, analytical procedure is as follows:
1) lipid acid extracts:
Getting the volume that 50mg or 100mg freeze-dried algae powder is placed on tool Telfnon bottle screw cap is in the phial of 15-20ml; place a little magnetic bar again; add 2-4ml10%DMSO-Methanol solution, 40 DEG C of sand baths (beaker containing sand is placed on constant-temperature heating magnetic stirring apparatus) 5 minutes; Then stir extracting 30 minutes at 4 DEG C of lower magnetic forces, 3500 leave the heart, and transfer supernatant liquor is in another bottle.Remaining algae-residue adds the ether of 1:1 again, normal hexane 4-8ml4 DEG C lower magnetic force stirs extracting 1 hour, and 3500 leave the heart, and transfer supernatant liquor is in an above-mentioned bottle.Said process can be repeated until algae-residue bleaches.In above-mentioned merging extract, add pure water makes four (water, DMSO-Methanol, ether, normal hexane) volume ratio be 1:1:1:1, concussion phase-splitting, pipetting organic phase transfers in another phial, blow to one-tenth concentrated solution with nitrogen in stink cupboard, then transfer in 1.5ml plastic centrifuge tube weighed in advance, then dry up to constant weight with nitrogen.
2) fatty acid analysis:
After method is extracted above, with n-hexane dissolution, (chromatographic condition is carrier gas: nitrogen flow 1ml/min, hydrogen flowing quantity 30ml/min, air flow quantity 300ml/min to use Agilent6820 gas chromatograph to carry out gas chromatographic analysis, injector temperature: 280 DEG C, detector temperature: 280 DEG C, detector type: FID, chromatographic column: DB-5 capillary chromatographic column (30m × 0.25mm, 0.25 μm), splitting ratio: 4:1.Analytical procedure: marker method (gas-chromatography nitrogen does carrier gas, is equivalent to the moving phase of liquid chromatography).
As shown in Figure 2, increase temperature rear wild type control strain ENN11 growth and obviously slow down, be cultured to the 10th day wild type control strain ENN11 and turn yellow dead, and though mutant strain ENN11-3 growth is also affected, but slowly can also grow, there is no death, show obvious high temperature resistant advantage.When being cultured to the 9th day, the fatty acid content of mutant strain ENN11-3 is compared wild strain with EPA content and is all improved to some extent.Wild contrast strain is dead afterwards, is under some influence, can also effectively accumulates although the fatty acid content of mutant strain ENN11-3 and EPA content accumulate under High Temperature Stress.When cultivating the 13rd day, the total fatty acid content of mutant strain ENN11-3 and EPA content reach the highest, within the 15th day, decline (Fig. 3) to some extent.
The ENN11-3 13d of table 2 gas Chromatographic Determination collects total fat component of sample:
Fatty acid component analysis shows, and its fat content is up to 41.1%, and wherein about 75.5% is C16-C18 lipid acid, 65.3% is C16 lipid acid, 10.2% is C18 lipid acid, and unsaturated fatty acids accounts for 37.4% of fatty acid total amount, can as the raw materials for production of biofuel.As can be seen here, algae strain ENN11-3 of the present invention can be used for production of biodiesel.
The unsaturated fatty acids of ENN11-3 accounts for 37.4% of fatty acid total amount, and content of polyunsaturated fatty acid accounts for 12.7% of fatty acid total amount, and wherein EPA content accounts for 4.5% of fatty acid total amount, and unsaturated fatty acid content enriches, and can be used for the production of polyunsaturated fatty acid.
The outdoor tubular type high temperature cultivation of embodiment 2ENN11-3 algae strain
Be seeded in 60*5cm tubular reactor by the algae kind being in logarithmic phase, use sea water medium to cultivate, compare with wild-type ENN11, every algae strain arranges two Duplicate Samples, is placed in the natural lighting of outdoor Arrangement for heating or cooling water in receptacle for live fish temperature control and carries out high temperature cultivation.Between incubation period, condition is as follows: cultivate first 5 days for natural temperature, within 24 hours, range of temperature is mainly between 17-37 DEG C.Be cultured to the 6th day to start to increase temperature, day temperature multidimensional is held in about 40 DEG C, and nocturnal temperature substantially also maintains more than 25 DEG C (Fig. 4).The illumination variation scope sunny side of 9:00-17:00 is mainly between 300-800umol/m2/s, and shady face is less than 200umol/m2/s substantially.Pass into a certain proportion of CO2 (purity more than 95%), control ph is between 7-9.Sampling and measuring dry weight (Fig. 5) is interrupted in culturing process.
Be cultured to the 10th day, because pyroprocessing pressure is excessive, two strain algae strains all turn yellow death.Cultivate first 5 days two strain algae dry weights to be more or less the same, increasing temperature from the 6th day, the biomass of mutant strain ENN11-3 is apparently higher than wild strain ENN11.The 5-9 days increased temperature, the average dry weight rate of growth of mutant strain ENN11-3 is 0.345g/ (L*d), and wild strain ENN11 is 0.131g/ (L*d), the mutant strain speed of growth is at high temperature 2.63 times of wild strain, shows stronger heat-resisting ability.
Embodiment 3 ENN11-3 intends indoor culture and the oil and fat accumulation of Nannochloropsis oceanica
The board-like high temperature cultivation in outdoor of embodiment 3 ENN11-3 algae strain
The algae kind being in logarithmic phase is seeded in 50*50*5cm plate-type reactor, sea water medium is used to cultivate, compare with wild-type ENN11, be placed in summer outdoor natural high-temperature condition and carry out grown cultures, between incubation period, condition is as follows: culture cycle is 11 days, the range of temperature of 9:00-17:00 is mainly between 35-40 DEG C, and nocturnal temperature is many at about 25 DEG C (Fig. 6).The illumination variation scope sunny side of 9:00-17:00 is mainly between 500-1000umol/m2/s, and shady face light intensity is substantially at about 300umol/m2/s.Pass into a certain proportion of CO2 (purity more than 95%), control ph is between 7-9.Sampling and measuring dry weight (Fig. 7) is interrupted in culturing process.Cultivate and within the 8th day, collect part algae liquid, obtain algae mud by method that is centrifugal or natural subsidence, algae mud is carried out vacuum lyophilization again, gets appropriate algae powder and carry out gas chromatographic analysis, method is with embodiment 1, and lipid acid content in algae powder is shown in Fig. 8.
The highest multidimensional of outdoor board-like high temperature cultivation algae liquid temp is held in about 35 DEG C, and be up to 40 DEG C, under this temperature condition, mutant strain ENN11-3 is obvious compared with the advantage of wild-type ENN11.The average dry weight rate of growth of mutant strain ENN11-3 is 0.272g/ (L*d), and wild strain ENN11 is 0.221g/ (L*d), and the board-like average growth rate of ENN11-3 improves 23.1% than ENN11.The EPA percentage composition cultivating 8 days ENN11-3 is 4.08%, and total fat percentage composition is 12.66%, and wild strain ENN11 is respectively 3.58% and 11.99% by contrast.
Do not have the accumulation of implementing measure induced lipolysis acid in the present embodiment, as taked corresponding measure, fatty acid content can reach higher level.
Embodiment 4 mutant strain ENN11-3 height is containing CO
2waste gas cultivation
The algae kind being in logarithmic phase is seeded in 50*50*5cm plate-type reactor, sea water medium is used to carry out cultivating (not adding C source), be placed in outdoor natural condition and carry out grown cultures, between incubation period, natural condition are as follows: in experimentation, 9:00-17:00 temperature multidimensional is held between 30-35 DEG C, and night, multidimensional was held in about 20 DEG C (Fig. 9).9:00-17:00 intensity of illumination variation range sunny side is mainly at 500-1000 μm of ol/m
2/ s, shady face light intensity is substantially at 300 μm of ol/m
2about/s.Pass into a certain proportion of CO
2(chemical plant waste gas, purity more than 95%), controls pH value between 6.9-8.1.Sampling and measuring biomass (Figure 10) is interrupted in culturing process.Cultivate and within the 7th day, collect part algae liquid, obtain algae mud by method that is centrifugal or natural subsidence, algae mud is being carried out vacuum lyophilization, is getting appropriate algae powder and carry out gas chromatographic analysis, method is with embodiment 1, and lipid acid content in algae powder is shown in Figure 11.
Do not have the accumulation of implementing measure induced lipolysis acid in the present embodiment, cultivate 7 days fatty acid content and reach 15.462%, EPA content reaches 3.611%.As taked corresponding measure, fatty acid content can be higher.
Use waste gas to cultivate 13 days in the present embodiment, mutant strain ENN11-3 breeding situations is good.In 0-9 days cultured continuously, output on average can reach 20.7g/m
2/ d.C source is not added in substratum during microdisk electrode, the CO that frond growth is fixing in the present embodiment
2from a certain proportion of CO passed in culturing process
2(CO
2derive from chemical plant, purity more than 95%), about can fix 2 tons of CO with algae per ton
2(yellow wise, Wang Weiliang, Li Yuanguang, Xie Jingli, model Jian Hua, Tao Liming. the thinking of development of micro-algae energy technology exploitation and industrialization and strategy, biotechnology journal, 2010,26 (7): 907-913), in the present embodiment, mutant strain ENN11-3 can realize CO
2reduce discharging 41.4g/m
2/ d.
embodiment 5 mutant strain ENN11-3 is used for the production of nutritious prod, feed, bait
Protein content in algae strain provided by the invention is 20 ~ 30%, and lipid content is 40 ~ 60%, and wherein the content of unsaturated fatty acids accounts for the 35%-50% of total fatty acid content, is the optimum feed stock source of cooking nutritious prod, feed, bait.
Applicant states, the present invention illustrates detailed features of the present invention and method by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and method, does not namely mean that the present invention must rely on above-mentioned detailed features and method could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the increase of accessory, the concrete way choice etc. of condition selected by the present invention, all drops within protection scope of the present invention and open scope.
Claims (17)
1. intend Nannochloropsis oceanica ENN11-3 (Nannochloropsis sp.) for one kind, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation date is on September 5th, 2012, and deposit number is CGMCC No.6446.
2. the as claimed in claim 1 cultural method intending Nannochloropsis oceanica ENN11-3, described method comprises and is seeded in sea water medium by plan Nannochloropsis oceanica algae strain ENN11-3 green cell, wherein intensity of illumination 50-500 μm of ol/m
2.s; PH value 7-9; Temperature 25-42 DEG C; And in incubation period, by passing into the air that carbonic acid gas volume fraction is 1.5-2% in nutrient solution.
3. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 2, it is characterized in that, described intensity of illumination is 100-450 μm of ol/m
2.s.
4. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 3, it is characterized in that, described intensity of illumination is 150-400 μm of ol/m
2.s.
5. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 4, it is characterized in that, described intensity of illumination is 200-350 μm of ol/m
2.s.
6. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 5, it is characterized in that, described intensity of illumination is 250-300 μm of ol/m
2.s.
7. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 2, it is characterized in that, pH value is 7.5-8.5.
8. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 7, it is characterized in that, pH value is 8.
9. the cultural method intending Nannochloropsis oceanica ENN11-3 as claimed in claim 2, it is characterized in that, temperature is 30-38 DEG C.
10. the application of plan Nannochloropsis oceanica ENN11-3 as claimed in claim 1 in lipid acid is produced.
11. apply as claimed in claim 10, it is characterized in that, described lipid acid is capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid.
12. intend the application of Nannochloropsis oceanica ENN11-3 in biofuel is produced as claimed in claim 1.
13. intend the application of Nannochloropsis oceanica ENN11-3 in environment protection as claimed in claim 1.
14. apply as claimed in claim 13, it is characterized in that, described environment protection is CO
2reduce discharging.
15. intend Nannochloropsis oceanica ENN11-3 is as claimed in claim 1 intending the application in the cultivation of Nannochloropsis oceanica high temperature, and wherein, described high temperature is 35-42 DEG C.
16. intend Nannochloropsis oceanica ENN11-3 is as claimed in claim 1 preparing the application comprised in the medicine of capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid.
17. plan Nannochloropsis oceanica ENN11-3 as claimed in claim 1 are comprising the purposes in the food of capric acid, lauric acid, myristic acid, palmitinic acid, Zoomeric acid, stearic acid, octadecenoic acid, octadecadienoic acid, gamma-linolenic acid, arachidonic acid and/or timnodonic acid, healthcare products, feed and/or the production of fishery products bait.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN104293676B (en) * | 2014-09-09 | 2017-09-15 | 新奥科技发展有限公司 | One plant is intended Nannochloropsis oculata mutant strain and its application |
| CN104371929B (en) * | 2014-09-09 | 2017-08-25 | 新奥科技发展有限公司 | One plant is intended Nannochloropsis oculata mutant strain and its application |
| CN104560720B (en) * | 2014-12-12 | 2017-06-23 | 中国海洋大学 | Intend double spore algaes and its application and cultural method in one plant of ocean |
| CN106701586A (en) * | 2015-08-19 | 2017-05-24 | 中国科学院大连化学物理研究所 | Method for culturing EPA-rich microalgae by using alkaline lake brine |
| CN106260630A (en) * | 2016-08-08 | 2017-01-04 | 新奥科技发展有限公司 | A kind of improve the pig feed of unsaturated content of fatty acid in pork fat |
| CN107375345A (en) * | 2017-08-25 | 2017-11-24 | 谱尼测试集团股份有限公司 | Intend Nannochloropsis oculata water extract and its radioresistance purposes |
| CN110604234A (en) * | 2019-05-22 | 2019-12-24 | 中国科学院水生生物研究所 | Application of Chlorella sorokiniana in functional feed for improving health condition of fishes |
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